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- Creators: Hamilton, Robert, 1937-
- Creators: Arizona State University
Biological and immunological characterization of plant-produced HIV-1 Gag/dgp41 virus-like particles
Description
Anti-retroviral drugs and AIDS prevention programs have helped to decrease the rate of new HIV-1 infections in some communities, however, a prophylactic vaccine is still needed to control the epidemic world-wide. Despite over two decades of research, a vaccine against HIV-1 remains elusive, although recent clinical trials have shown promising results. Recent successes have focused on highly conserved, mucosally-targeted antigens within HIV-1 such as the membrane proximal external region (MPER) of the envelope protein, gp41. MPER has been shown to play critical roles in the viral mucosal transmission, though this peptide is not immunogenic on its own. Gag is a structural protein configuring the enveloped virus particles, and has been suggested to constitute a target of the cellular immunity potentially controlling the viral load. It was hypothesized that HIV-1 enveloped virus-like particles (VLPs) consisting of Gag and a deconstructed form of gp41 comprising the MPER, transmembrane, and cytoplasmic domains (dgp41) could be expressed in plants. Plant-optimized HIV-1 genes were constructed and expressed in Nicotiana benthamiana by stable transformation, or transiently using a tobacco mosaic virus-based expression system or a combination of both. Results of biophysical, biochemical and electron microscopy characterization demonstrated that plant cells could support not only the formation of HIV-1 Gag VLPs, but also the accumulation of VLPs that incorporated dgp41. These particles were purified and utilized in mice immunization experiments. Prime-boost strategies combining systemic and mucosal priming with systemic boosting using two different vaccine candidates (VLPs and CTB-MPR - a fusion of MPER and the B-subunit of cholera toxin) were administered to BALB/c mice. Serum antibody responses against both the Gag and gp41 antigens could be elicited in mice systemically primed with VLPs and these responses could be recalled following systemic boosting with VLPs. In addition, mucosal priming with VLPs allowed for a robust boosting response against Gag and gp41 when boosted with either candidate. Functional assays of these antibodies are in progress to test the antibodies' effectiveness in neutralizing and preventing mucosal transmission of HIV-1. This immunogenicity of plant-based Gag/dgp41 VLPs represents an important milestone on the road towards a broadly-efficacious and inexpensive subunit vaccine against HIV-1.
ContributorsKessans, Sarah (Author) / Mor, Tsafrir S (Thesis advisor) / Matoba, Nobuyuki (Committee member) / Mason, Hugh (Committee member) / Hogue, Brenda (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2011
Description
Yannis Constantinidis was the last of the handful of composers referred to collectively as the Greek National School. The members of this group strove to create a distinctive national style for Greece, founded upon a synthesis of Western compositional idioms with melodic, rhyhmic, and modal features of their local folk traditions. Constantinidis particularly looked to the folk melodies of his native Asia Minor and the nearby Dodecanese Islands. His musical output includes operettas, musical comedies, orchestral works, chamber and vocal music, and much piano music, all of which draws upon folk repertories for thematic material. The present essay examines how he incorporates this thematic material in his piano compositions, written between 1943 and 1971, with a special focus on the 22 Songs and Dances from the Dodecanese. In general, Constantinidis's pianistic style is expressed through miniature pieces in which the folk tunes are presented mostly intact, but embedded in accompaniment based in early twentieth-century modal harmony. Following the dictates of the founding members of the Greek National School, Manolis Kalomiris and Georgios Lambelet, the modal basis of his harmonic vocabulary is firmly rooted in the characteristics of the most common modes of Greek folk music. A close study of his 22 Songs and Dances from the Dodecanese not only offers a valuable insight into his harmonic imagination, but also demonstrates how he subtly adapts his source melodies. This work also reveals his care in creating a musical expression of the words of the original folk songs, even in purely instrumental compositon.
ContributorsSavvidou, Dina (Author) / Hamilton, Robert (Thesis advisor) / Little, Bliss (Committee member) / Meir, Baruch (Committee member) / Thompson, Janice M (Committee member) / Arizona State University (Publisher)
Created2011
Description
This paper describes six representative works by twentieth-century Chinese composers: Jian-Zhong Wang, Er-Yao Lin, Yi-Qiang Sun, Pei-Xun Chen, Ying-Hai Li, and Yi Chen, which are recorded by the author on the CD. The six pieces selected for the CD all exemplify traits of Nationalism, with or without Western influences. Of the six works on the CD, two are transcriptions of the Han Chinese folk-like songs, one is a composition in the style of the Uyghur folk music, two are transcriptions of traditional Chinese instrumental music dating back to the eighteenth century, and one is an original composition in a contemporary style using folk materials. Two of the composers, who studied in the United States, were strongly influenced by Western compositional style. The other four, who did not study abroad, retained traditional Chinese style in their compositions. The pianistic level of difficulty in these six pieces varies from intermediate to advanced level. This paper includes biographical information for the six composers, background information on the compositions, and a brief analysis of each work. The author was exposed to these six pieces growing up, always believing that they are beautiful and deserve to be appreciated. When the author came to the United States for her studies, she realized that Chinese compositions, including these six pieces, were not sufficiently known to her peers. This recording and paper are offered in the hopes of promoting a wider familiarity with Chinese music and culture.
ContributorsLuo, Yali, D.M.A (Author) / Hamilton, Robert (Thesis advisor) / Campbell, Andrew (Committee member) / Pagano, Caio (Committee member) / Cosand, Walter (Committee member) / Rogers, Rodney (Committee member) / Arizona State University (Publisher)
Created2012
Description
The purpose of this project was to examine the lives and solo piano works of four members of the early generation of female composers in Taiwan. These four women were born between 1950 and 1960, began to appear on the Taiwanese musical scene after 1980, and were still active as composers at the time of this study. They include Fan-Ling Su (b. 1955), Hwei-Lee Chang (b. 1956), Shyh-Ji Pan-Chew (b. 1957), and Kwang-I Ying (b. 1960). Detailed biographical information on the four composers is presented and discussed. In addition, the musical form and features of all solo piano works at all levels by the four composers are analyzed, and the musical characteristics of each composer's work are discussed. The biography of a fifth composer, Wei-Ho Dai (b. 1950), is also discussed but is placed in the Appendices because her piano music could not be located. This research paper is presented in six chapters: (1) Prologue; the life and music of (2) Fan-Ling Su, (3) Hwei-Lee Chang, (4) Shyh-Ji Pan-Chew, and (5) Kwang-I Ying; and (6) Conclusion. The Prologue provides an overview of the development of Western classical music in Taiwan, a review of extant literature on the selected composers and their music, and the development of piano music in Taiwan. The Conclusion is comprised of comparisons of the four composers' music, including their personal interests and preferences as exhibited in their music. For example, all of the composers have used atonality in their music. Two of the composers, Fan-Ling Su and Kwang-I Ying, openly apply Chinese elements in their piano works, while Hwei-Lee Chang tries to avoid direct use of the Chinese pentatonic scale. The piano works of Hwei-Lee Chang and Shyh-Ji Pan-Chew are chromatic and atonal, and show an economical usage of material. Biographical information on Wei-Ho Dai and an overview of Taiwanese history are presented in the Appendices.
ContributorsWang, Jinding (Author) / Pagano, Caio (Thesis advisor) / Campbell, Andrew (Committee member) / Humphreys, Jere T. (Committee member) / Meyer-Thompson, Janice (Committee member) / Norton, Kay (Committee member) / Arizona State University (Publisher)
Created2011
Description
CTB-MPR649-684 is a translational fusion protein consisting of the cholera toxin B subunit (CTB) and the conserved residues 649-684 of gp41 membrane proximal region (MPR). It is a candidate vaccine component aimed at early steps of the HIV-1 infection by blocking viral mucosal transmission. Bacterially produced CTB-MPR was previously shown to induce HIV-1 transcytosis-blocking antibodies in mice and rabbits. However, the induction of high-titer MPR specific antibodies with HIV-1 transcytosis blocking ability remains a challenge as the immuno-dominance of CTB overshadows the response to MPR. X-ray crystallography was used to investigate the structure of CTB-MPR with the goal of identifying potential solutions to improve the immune response of MPR. Various CTB-MPR variants were designed using different linkers connecting the two fusion proteins. The procedures for over-expression E. coli and purification have been optimized for each of the variants of CTB-MPR. The purity and oligomeric homogeneity of the fusion protein was demonstrated by electrophoresis, size-exclusion chromatography, dynamic light scattering, and immuno-blot analysis. Crystallization conditions for macroscopic and micro
ano-crystals have been established for the different variants of the fusion protein. Diffraction patterns were collected by using both conventional and serial femto-second crystallography techniques. The two crystallography techniques showed very interesting differences in both the crystal packing and unit cell dimensions of the same CTB-MPR construct. Although information has been gathered on CTB-MPR, the intact structure of fusion protein was not solved as the MPR region showed only weak electron density or was cleaved during crystallization of macroscopic crystals. The MPR region is present in micro
ano-crystals, but due to the severe limitation of the Free Electron Laser beamtime, only a partial data set was obtained and is insufficient for structure determination. However, the work of this thesis has established methods to purify large quantities of CTB-MPR and has established procedures to grow crystals for X-ray structure analysis. This has set the foundation for future structure determination experiments as well as immunization studies.
ano-crystals have been established for the different variants of the fusion protein. Diffraction patterns were collected by using both conventional and serial femto-second crystallography techniques. The two crystallography techniques showed very interesting differences in both the crystal packing and unit cell dimensions of the same CTB-MPR construct. Although information has been gathered on CTB-MPR, the intact structure of fusion protein was not solved as the MPR region showed only weak electron density or was cleaved during crystallization of macroscopic crystals. The MPR region is present in micro
ano-crystals, but due to the severe limitation of the Free Electron Laser beamtime, only a partial data set was obtained and is insufficient for structure determination. However, the work of this thesis has established methods to purify large quantities of CTB-MPR and has established procedures to grow crystals for X-ray structure analysis. This has set the foundation for future structure determination experiments as well as immunization studies.
ContributorsLee, Ho-Hsien (Author) / Fromme, Petra (Thesis advisor) / Mor, Tsafrir (Committee member) / Ros, Alexandra (Committee member) / Arizona State University (Publisher)
Created2015
Description
The HIV-1 pandemic continues to cause millions of new infections and AIDS-related deaths each year, and a majority of these occur in regions of the world with limited access to antiretroviral therapy. Therefore, an HIV-1 vaccine is still desperately needed. The most successful HIV-1 clinical trial to date used a non-replicating canarypox viral vector and protein boosting, yet its modest efficacy left room for improvement. Efforts to derive novel vectors which can be both safe and immunogenic, have spawned a new era of live, viral vectors. One such vaccinia virus vector, NYVAC-KC, was specifically designed to replicate in humans and had several immune modulators deleted to improve immunogenicity and reduce pathogenicity. Two NYVAC-KC vectors were generated: one expressing the Gag capsid, and one with deconstructed-gp41 (dgp41), which contains an important neutralizing antibody target, the membrane proximal external region (MPER). These vectors were combined with HIV-1 Gag/dgp41 virus-like particles (VLPs) produced in the tobacco-relative Nicotiana benthamiana. Different plant expression vectors were compared in an effort to improve yield. A Geminivirus-based vector was shown to increase the amount of MPER present in VLPs, thus potentially enhancing immunogenicity. Furthermore, these VLPs were shown to interact with the innate immune system through Toll-like receptor (TLR) signaling, which activated antigen presenting cells to induce a Th2-biased response in a TLR-dependent manner. Furthermore, expression of Gag and dgp41 in NYVAC-KC vectors resulted in activation of antiviral signaling pathways reliant on TBK1/IRF3, which necessitated the use of higher doses in mice to match the immunogenicity of wild-type viral vectors. VLPs and NYVAC-KC vectors were tested in mice, ultimately showing that the best antibody and Gag-specific T cell responses were generated when both components were administered simultaneously. Thus, plant-produced VLPs and poxvirus vectors represent a highly immunogenic HIV-1 vaccine candidate that warrants further study.
ContributorsMeador, Lydia Rebecca (Author) / Mor, Tsafrir S (Thesis advisor) / Jacobs, Bertram L (Thesis advisor) / Blattman, Joseph N (Committee member) / Mason, Hugh S (Committee member) / Arizona State University (Publisher)
Created2016
ContributorsCarpenter, Ellon (Performer) / Cosand, Walter, 1950- (Performer) / Tescarollo, Hamilton (Performer) / Hamilton, Robert, 1937- (Performer) / ASU Library. Music Library (Publisher)
Created2006-04-12
ContributorsHamilton, Robert, 1937- (Performer) / ASU Library. Music Library (Publisher)
Created1987-09-13
ContributorsHamilton, Robert, 1937- (Performer) / ASU Library. Music Library (Publisher)
Created2007-10-07
ContributorsHamilton, Robert, 1937- (Performer) / ASU Library. Music Library (Publisher)
Created1988-06-18